Polynucleotides encoding alpha-galactosidase a for the treatment of fabry disease

Abstract

The invention relates to mRNA therapy for the treatment of Fabry disease. mRNAs for use in the invention, when administered in vivo, encode human the α-galactosidase A (GLA), isoforms thereof, functional fragments thereof, and fusion proteins comprising GLA. mRNAs of the invention are preferably encapsulated in lipid nanoparticles (LNPs) to effect efficient delivery to cells and/or tissues in subjects, when administered thereto. mRNA therapies of the invention increase and/or restore deficient levels of GLA expression and/or activity in subjects. mRNA therapies of the invention further decrease levels of toxic metabolites associated with deficient GLA activity in subjects, namely Gb3 and lyso-Gb3.

What is claimed is: 
     
         1 . A method of treating Fabry disease in a subject, comprising administering a composition comprising a messenger RNA (mRNA) and a pharmaceutically acceptable carrier, wherein the mRNA comprises an open reading frame (ORF) encoding a human α-galactosidase A (GLA) polypeptide of SEQ ID NO: 1, and wherein the ORF comprises a nucleotide sequence at least 95% identical to SEQ ID NO: 79. 
     
     
         2 . The method of  claim 1 , wherein the nucleotide sequence is at least 96%, at least 97% or 100% identical to SEQ ID NO: 79. 
     
     
         3 . The method of  claim 1 , wherein the mRNA comprises a 5′ terminal cap, and a poly-A tail about 100 residues in length. 
     
     
         4 . The method of  claim 1 , wherein at least 95%, at least 99%, or 100% of uridines in the ORF are modified uridines. 
     
     
         5 . The method of  claim 4 , wherein the modified uridines are 5-methoxyuridines. 
     
     
         6 . The method of  claim 1 , wherein treatment reduces Gb3 or lyso-Gb3 in plasma or tissues of the subject. 
     
     
         7 . The method of  claim 1 , wherein the mRNA is intravenously administered. 
     
     
         8 . A method of treating Fabry disease in a subject, comprising administering a lipid nanoparticle (LNP) comprising an mRNA and a compound having the formula: 
       
         
           
           
               
               
           
         
         wherein the mRNA comprises an ORF encoding a human GLA polypeptide of SEQ ID NO: 1. 
       
     
     
         9 . The method of  claim 8 , wherein the LNP comprises a phospholipid, a structural lipid, and a PEG lipid. 
     
     
         10 . The method of  claim 8 , wherein the ORF comprises a nucleotide sequence at least 95%, at least 96%, at least 97% or 100% identical to SEQ ID NO: 79. 
     
     
         11 . The method of  claim 8 , wherein at least 95%, at least 99% or 100% of uridines in the ORF are modified uridines. 
     
     
         12 . The method of  claim 11 , wherein the modified uridines are 5-methoxyuridines. 
     
     
         13 . The method of  claim 8 , wherein treatment reduces Gb3 or lyso-Gb3 in plasma or tissues of the subject. 
     
     
         14 . The method of  claim 8 , wherein the LNP is intravenously administered. 
     
     
         15 . A method of treating Fabry disease in a subject, comprising administering an LNP comprising (i) an mRNA comprising an ORF encoding a human GLA polypeptide, wherein the mRNA comprises a nucleotide sequence at least 95% identical to SEQ ID NO: 79, and (ii) an ionizable lipid, wherein the ionizable lipid is a compound selected from Compound 1 to Compound 232, salts and stereoisomers thereof, and combinations thereof. 
     
     
         16 . The method of  claim 15 , wherein the compound is selected from Compound 1 to Compound 147, salts and stereoisomers thereof, and combinations thereof. 
     
     
         17 . The method of  claim 15 , wherein the compound is Compound 18, a salt or a stereoisomer thereof, or any combination thereof. 
     
     
         18 . The method of  claim 17 , wherein the LNP comprises a phospholipid, a PEG-lipid, and a structural lipid. 
     
     
         19 . The method of  claim 15 , wherein the LNP comprises from about 45 mol % to about 55 mol % of ionizable lipid. 
     
     
         20 . The method of  claim 15 , wherein the LNP comprises from about 1 mol % to about 20 mol % of phospholipid. 
     
     
         21 . The method of  claim 15 , wherein the LNP comprises from about 35 mol % to about 40 mol % of structural lipid. 
     
     
         22 . The method of  claim 15 , wherein the LNP comprises from about 2 mol % to about 4 mol % of PEG lipid. 
     
     
         23 . The method of  claim 17 , wherein the LNP comprises about 45 mol % to about 55 mol % of Compound 18. 
     
     
         24 . The method of  claim 18 , wherein the LNP comprises from about 1 mol % to about 20 mol % of phospholipid. 
     
     
         25 . The method of  claim 18 , wherein the LNP comprises from about 35 mol % to about 40 mol % of structural lipid. 
     
     
         26 . The method of  claim 18 , wherein the LNP comprises from about 2 mol % to about 4 mol % of PEG lipid. 
     
     
         27 . The method of  claim 15 , wherein the nucleotide sequence is at least 96%, at least 97% or 100% identical to SEQ ID NO: 79. 
     
     
         28 . The method of  claim 15 , wherein at least 95%, at least 99% or 100% of are modified uridines. 
     
     
         29 . The method of  claim 28 , wherein the modified uridines are 5-methoxyuridines. 
     
     
         30 . The method of  claim 18 , wherein the nucleotide sequence is at least 96%, at least 97% or 100% identical to SEQ ID NO: 79. 
     
     
         31 . The method of  claim 18 , wherein at least 95%, at least 99% or 100% of are modified uridines. 
     
     
         32 . The method of  claim 31 , wherein the modified uridines are 5-methoxyuridines. 
     
     
         33 . The method of  claim 23 , wherein the nucleotide sequence is at least 96%, at least 97% or 100% identical to SEQ ID NO: 79. 
     
     
         34 . The method of  claim 23 , wherein at least 95%, at least 99% or 100% of are modified uridines. 
     
     
         35 . The method of  claim 34 , wherein the modified uridines are 5-methoxyuridines. 
     
     
         36 . The method of  claim 15 , wherein treatment reduces Gb3 or lyso-Gb3 in plasma or tissues of the subject. 
     
     
         37 . The method of  claim 15 , wherein the LNP is intravenously administered.